Sie sind auf Seite 1von 43

Tools of the Trade

Laboratory Notebook

Objectives of a Good Lab Notebook


(a) State what was done
(b) State what was observed
(c) Be easily understandable to someone
else
Tools of the Trade
Laboratory Notebook

Bad Laboratory Practice (A Recent Legal Case)


Medichem Pharmaceuticals v. Rolabo Pharmaceuticals

Two Patents describe a method for making the antihistamine drug Loratidine (Claritin)
- US sales of $2.7 billion
- the two patents are essentially identical
- Medichem sued to invalidate Rolabo patent and claimed priority
- Medichem had to prove it used the method to make loratidine before Rolabo did

A co-inventor’s lab notebook was a primary piece of evidence to support Medichem’s claim
- documented analysis of a sample claimed to be made using the patented method
- NMR spectral data confirmed the production of loratidine

The evidence was not enough to support Medichem's claim of reduction to practice
- NMR data do not show the process by which loratidine was made
- lab books were not witnessed

Rolabo Pharmaceuticals won the case (and the rights to make Loratidine)
because of problems with a Lab Notebook!!
Nature Reviews Drug Discovery (2006) 5, 180
Tools of the Trade
Weight Measurements

1.) Analytical Balance (principal of operation):


(i) sample on balance pushes the pan down with a force equal to m x g
 M is mass of object
 g is acceleration of gravity
(ii) balance pan with equal and opposing mass
 Mechanical – standard masses
 Electronic – opposing electromagnetic force
(iii) tare –mass of empty vessel (pan)

l1 l2

Double-pan balance
(a) balance beam suspended on a
sharp knife edge
(b) Standard weights are added to
m1 m2 the second pan to balance
sample weight
m m (c) Weight of sample is equal to the
total weight of standards
m1l1 = m2l2
Tools of the Trade
Weight Measurements

Single-pan balance
(a) balance beam suspended on a sharp knife edge
(b) Sample pan is balanced by counterweights on right
(c) Knob adjusted to remove weights from a bar above the pan
(d) Pan is moved back to its original position and the removed
weights equals the mass of the sample.
Tools of the Trade
Weight Measurements

Electronic balance
(a) Uses electromagnetic force to return the pan to original position
(b) Electric current required to generate the force is proportional to sample mass

Determines amount of deflection


of pan due to mass of sample

Increase in current causes


magnetic field that raises pan
Tools of the Trade
Weight Measurements

2.) Methods of Weighing:


(i) Basic operational rules
 Chemicals should never be placed directly on the weighing pan
- corrode and damage the pan may affect accuracy
- not able to recover all of the sample
 Balance should be in arrested position when load/unload pan
 Half-arrested position when dialing weights
- dull knife edge and decrease balance sensitivity  accuracy
(ii) Weight by difference:
 Useful for samples that change weight upon exposure to the
atmosphere
- hygroscopic samples (readily absorb water from the air)

Weight of sample = ( weight of sample + weight of container) – weight of container

(iii) Taring:
 Done on many modern electronic balances
 Container is set on balance before sample is added
 Container’s weight is set automatically to read “0”
Tools of the Trade
Weight Measurements

3.) Errors in Weighing: Sources


(i) Any factor that will change the apparent mass of the sample
 Dirty or moist sample container
- also may contaminate sample
 Sample not at room temperature Office dust
- avoid convection air currents (push/lift pan)
 Adsorption of water, etc. from air by sample
 Vibrations or wind currents around balance
 Non-level balance
Tolerance (mg) Tolerance (mg)
Grams Class 1 Class 2 Milligrams Class 1 Class 2
500 1.2 2.5 500 0.010 0.025
200 0.5 1.0 200 0.010 0.025
100 0.25 0.50 100 0.010 0.025
50 0.12 0.25 50 0.010 0.014
20 0.074 0.10 20 0.010 0.014
10 0.050 0.074 10 0.010 0.014
5 0.034 0.054 5 0.010 0.014
2 0.034 0.054 2 0.010 0.014
1 0.034 0.054 1 0.010 0.014
Tools of the Trade
Weight Measurements

3.) Errors in Weighing: Sources


(i) Any factor that will change the apparent mass of the sample
 Buoyancy errors – failure to correct for weight difference due to
displacement of air by the sample.

balsa Different displacement of


ice ice and balsa wood in water

 Correction for buoyancy to give true mass of sample


da
m' ( 1  )
dw
m
da
(1 )
m = true mass of sample d
m’ = mass read from balance
d = density of sample
da = density of air (0.0012 g/ml at 1 atm & 25oC)
dw = density of calibration weights (~ 8.0 g/ml)
Tools of the Trade
Weight Measurements

3.) Errors in Weighing: Sources


Example: The densities (g/ml) of several substances are:

acetic acid 1.05 CCl4 1.59 Sulfur 2.07


lithium 0.53 mercury 13.5 PbO2 9.4
lead 11.4 iridium 22.5

From the following figure:

predict which substance will have the smallest percentage buoyancy


correction and which will have the greatest.
Tools of the Trade
Weight Measurements

3.) Errors in Weighing: Sources


(i) Any factor that will change the apparent mass of the sample
 Density of air changes with temperature and pressure
 To get da under non standard conditions
B  0.3783V
d a  0.46468 ( )
T
B = Barometer pressure (torr)
V = vapor pressure of water in the air (torr)
T = air temperature (K)
Tools of the Trade
Volume Measurements

1.) Burets
(i) Purpose: used to deliver multiple aliquots of a liquid in known volumes
Buret Smallest Tolerance (mL)
volume (ml) graduation (ml)
5 0.01 ± 0.01
10 0.05 or 0.02 ± 0.02
25 0.1 ± 0.03
50 0.1 ± 0.05
100 0.2 ± 0.10

(ii) Correct use of buret


 Read buret at the bottom of a concave meniscus

Meniscus at 9.68 mL
Tools of the Trade
Volume Measurements

1.) Burets
(iii) always read the buret at the same eyelevel as the liquid
 Avoids parallax errors
eyelevel View from above

15.46 mL 15.31 mL
1% error
(iv) Consistently read all levels versus a given position on the nearest mark
Tools of the Trade
Volume Measurements

1.) Burets
(v) Estimate the buret reading to the nearest 1/10 of a division
(vi) expel all air bubbles from the stopcock prior to use
(vii) rinse the buret with a solution 2-3x before filling the buret for a titration
(viii) Near the end of a titration, volume of 1 drop or less per delivery should be
used with the buret.
Tools of the Trade
Volume Measurements

2.) Volumetric Flasks


(i) Purpose: used to prepare a solution of a single known volume
(ii) Types of volumetric flasks
Flask Capacity (mL) Tolerance (mL)
1 ± 0.02
2 ± 0.02
5 ± 0.02
10 ± 0.02
25 ± 0.03
50 ± 0.05
100 ± 0.08
200 ± 0.10
250 ± 0.12
500 ± 0.20
1000 ± 0.30
2000 ± 0.50
Tools of the Trade
Volume Measurements

2.) Volumetric Flasks


(iii) Correct use of volumetric flask
 Add reagent or solution to flask and dissolve in volume of solvent less
than the total capacity of the flask
 Slowly add more solvent until the meniscus bottom is level with the
calibration line.

stopper

 Stopper the flask and mix solution by inversion (40 or more times)
 (for later use) Remix by inverting the flask if the solution has been sitting
unused for more than several hours
 Glass adsorbs trace amount of chemicalsclean using acid wash
- adhere to surface
Tools of the Trade
Volume Measurements

3.) Pipets and Syringes


(i) Use to deliver a given volume of liquid
(ii) Types of pipets
 Transfer pipet
Transfer Pipet - similar to volumetric flask
Volume (mL) Tolerance (mL) - transfers a single volume  fill to calibration mark
0.5 ±0.006 - last drop does not drain out of the pipet  do not blow out
1 ±0.006 - more accurate than measuring pipet
2 ±0.006
3 ±0.01
4 ±0.01
5 ±0.01
 Measuring pipet
10 ±0.02 - calibrated similar to buret
15 ±0.03 - use to delivery a variable volume
20 ±0.03
25 ±0.03
50 ±0.05
100 ±0.08
Tools of the Trade
Volume Measurements

3.) Pipets and Syringes


(ii) Types of pipets
 Micropipet
- deliver volumes of 1 to 1000 mL (fixed & variable)
- uses disposable polypropylene tip
- stable in most aqueous and organic solvents (not chloroform)
- need periodic calibration
10% of pipet volume 100% of pipet volume

Pipet volume (mL) Accuracy (%) Precision (%) Accuracy (%) Precision (%)

Adjustable pipets

0.2-2 ±8 ±4 ±1.2 ±0.6

1-10 ±2.5 ±1.2 ±0.8 ±0.4

2.5-25 ±4.5 ±1.5 ±0.8 ±0.2

10-100 ±1.8 ±0.7 ±0.6 ±0.15

30-300 ±1.2 ±0.4 ±0.4 ±0.15

100-1000 ±1.6 ±0.5 ±0.3 ±0.12


Disposable tip
Fixed pipets

10 ±0.88 ±0.4

25 ±0.88 ±0.3

100 ±0.5 ±0.2

500 ±0.4 ±0.18

1000 ±0.33 ±0.12


Tools of the Trade
Volume Measurements

3.) Pipets and Syringes


(ii) Types of pipets
 Syringes
- deliver volumes of 1 to 500 mL
- accuracy & precision ~0.5-1%
- steel needle permits piercing stopper to transfer liquid under
controlled atmosphere
> attacked by strong acid and contaminate solution with iron

(iii) Correct use of pipets and syringes


 Use a bulb, never your mouth, for drawing solutions into pipets.
 Rinse pipets and syringes before using
- remove bubbles
Tools of the Trade
Filtration

1.) Mechanical separation of a liquid from the


undissolved particles floating in it.

2.) Purpose: used in gravimetric analysis for analysis of a


substance by the mass of a precipitate it produces
(i) Solid collected in paper or fritted-glass filters

3.) Process:
(i) pour slurry of precipitate down a glass rod to
prevent splattering.
(ii) dislodge solid from beaker/rod with rubber
policeman
(iii) use wash liquid (squirt bottle) to transfer particles to
filter paper
(iv) dry sample

Rubber policeman
Tools of the Trade
Drying

1.) Purpose: (i) to remove moisture from reagents or samples


(ii) to convert sample to a more readily analyzable form

2.) Oven Drying: commonly used for reagent or sample preparation


(i) Typically 110 oC for H2O removal
(ii) Use loose covers to prevent contamination from dust

3.) Dessicator: used to cool and store reagent or sample over long periods of
time.
(i) Contains a drying agent to absorb water from the atmosphere
(ii) Airtight seal
Experimental Error & Data Handling
Introduction

1.) There is error or uncertainty associated with every measurement.


(i) except simple counting

2.) To evaluate the validity of a measurement, it is necessary to evaluate its


error or uncertainty

You can read the name of


the boat on the left picture,
which is lost in the right
picture.

Can you read the tire


manufacturer?

Same Picture Different Levels of Resolution


Experimental Error & Data Handling
Significant Figures

1.) Definition: The minimum number of digits needed to write a given value
(in scientific notation) without loss of accuracy.
(i) Examples:

142.7 = 1.427 x 102


Both numbers have 4 significant figures
0.006302 = 6.302 x10-3

Zeros are simple place holders

2.) Zeros are counted as significant figures only if:


(i) occur between other digits in the number

9502.7 or 0.9907 Both zeros are significant figures

(ii) occur at the end of number and to the right of the decimal point

177.930 zero is a significant figure


Experimental Error & Data Handling
Significant Figures

3.) The last significant figure in any number is the first digit with any
uncertainty
(i) the minimum uncertainty is ± 1 unit in the last significant figure
(ii) if the uncertainty in the last significant figure is ≥ 10 units, then one less
significant figure should be used.
(iii) Example:

9.34 ± 0.02 3 significant figures


But

6.52 ± 0.12 should be 6.5 ± 0.1 2 significant figures

4.) Whenever taking a reading from an instrument, apparatus, graph, etc. always
estimate the result to the nearest tenth of a division
(i) avoids losing any significant figures in the reading process

7.45 cm
Experimental Error & Data Handling
Significant Figures

5.) Addition and Subtraction


(i) use the following procedure:
 Express all numbers using the same exponent
 Align all numbers with respect to the decimal point

1.25 x 105 12.5 x 104


2.48 x 104 2.48 x 104
+ 1.235 x 104 + 1.235 x 104

 Add or subtract using all given digits


 Round off the answer so that it has the same number of digits to
the right of the decimal as the number with the fewest decimal
places

12.5 x 104 1 decimal point


2.48 x 104
+ 1.235 x 104
16.215 x 104 = 16.2 x 104
Experimental Error & Data Handling
Significant Figures

5.) Addition and Subtraction


(i) use the following procedure:
 Round off the answer to the nearest digit in the least significant
figure.
 Consider all digits beyond the least significant figure when
rounding.
 If a number is exactly half-way between two digits, round to the
nearest even digit.
- minimizes round-off errors
 Examples:

3 sig. fig.: 12.534  12.5

4 sig. fig.: 11.126  11.13

4 sig. fig.: 101.250  101.2

3 sig. fig. 93.350  93.4


Experimental Error & Data Handling
Significant Figures

6.) Multiplication and Division


(i) use the following procedure:
 Express the answers in the same number of significant figures as
the number of digits in the number used in the calculation which
had the fewest significant figures.
 Examples:

3.261 x 10-5
x 1.78
3 significant figures
5.80 x 10-5

34.60
) 2.4287
4 significant figures
14.05
Experimental Error & Data Handling
Significant Figures

7.) Logarithms and Antilogarithms


(i) the logarithm of a number “a” is the value “b”, where:

a = 10b or Log(a) = b

(ii) example:
The logarithm of 100 is 2, since:
100 = 102

(iii) The antilogarithm of “b” is “a”


a = 10b

(iv) the logarithm of “a” is expressed in two parts

Log(339) = 2.530

character mantissa
Experimental Error & Data Handling
Significant Figures

7.) Logarithms and Antilogarithms


(v) when taking the logarithm of a number, the number of significant figures
in the resulting mantissa should be the same as the total number of
significant figures in the original number “a”

(vi) Example:

Log(5.403 x 10-8) = -7.2674

4 sig. fig. 4 sig. fig.

(vii) when taking the antilogarithm of a number, the number of significant


figures in the result should be the same as the total number of significant
figures in the mantissa of the original logarithm “b”

(viii) Example:

Antilog(-3.42) = 3.8 x 10-4

2 sig. fig. 2 sig. fig.


Experimental Error & Data Handling
Significant Figures

8.) Graphs
(i) use graph paper with enough rulings to accurately graph the results

(ii) plan the graph coordinates so that the data is spread over as much of the
graph as possible
(iii) in reading graphs, estimate values to the nearest 1/10 of a division on the
graph
Experimental Error & Data Handling
Significant Figures

8.) Graphs
(ii) plan the graph coordinates so that the data is spread over as much of the
graph as possible

(iii) in reading graphs, estimate values to the nearest 1/10 of a division on the
graph
Experimental Error & Data Handling
Errors

1.) Systematic (or Determinate) Error


(i) An error caused consistently in all results due to inappropriate methods or
experimental techniques.
(ii) Results in all measurements exhibiting a definite difference from the true
value.
(iii) This type of error can, in principal, be discovered and corrected.

Buret incorrectly calibrated


Experimental Error & Data Handling
Errors

2.) Random (or Indeterminate) Error


(i) An error caused by random variations in the measurement of a physical
quantity.
(ii) Results in a scatter of results centered on the true value for repeated
measurements on a single sample.
(iii) This type of error is always present and can never be totally eliminated

True value

Random Error Systematic Error


Experimental Error & Data Handling
Errors

3.) Accuracy and Precision


(i) Accuracy: refers to how close an answer is to the “true” value
 Generally, don’t know “true” value
 Accuracy is related to systematic error

(ii) Precision: refers to how the results of a single measurement compares


from one trial to the next
 Reproducibility
 Precision is related to random error

Low accuracy, low precision Low accuracy, high precision

High accuracy, low precision High accuracy, high precision


Experimental Error & Data Handling
Errors

4.) Absolute and Relative Uncertainty


(i) Both measures of the precision associated with a given measurement.
(ii) Absolute uncertainty: margin of uncertainty associated with a measurement
(iii) Example:
If a buret is calibrated to read within ± 0.02 mL, the absolute uncertainty
for measuring 12.35 mL is:

Absolute Uncertainty = 12.35 ± 0.02 mL


(iv) Relative uncertainty: compares the size of the absolute uncertainty with the
size of its associated measurement
Absolute Uncertaint y
Relative Uncertaint y 
Measured Value
(v) Example:
For a buret reading of 12.35 ± 0.02 mL, the relative uncertainty is:
(Make sure units cancel)
0.02 mL
Relative Uncertaint y(%)  (100)  0.16%  0.2%
12.35 mL 1 sig. fig.
Experimental Error & Data Handling
Errors

5.) Propagation of Uncertainty


(i) The absolute or relative uncertainty of a calculated result can be estimated
using the absolute or relative uncertainties of the values used to obtain that
result.
(ii) Addition and Subtraction
 The absolute uncertainty of a number calculated by addition or
subtraction is obtained by using the absolute uncertainties of
numbers used in the calculations as follows:

Abs . Uncert .Answer  Abs . Uncert .


value1    Abs . Uncert .
2
value 2 2


 Example: Value Abs. Uncert.


1.76 (± 0.03)
+ 1.89 (± 0.02)
- 0.59 (± 0.02)
Answer: 3.06

Abs . Uncert .Answer  0.032  0.022  0.022  0.04


Experimental Error & Data Handling
Errors

5.) Propagation of Uncertainty


(iii) Once the absolute uncertainty of the answer has been determined, its
relative uncertainty can also be calculated, as described previously.
 Example (using the previous example):

0.04
Re l . Uncert .(%)  ( 100 )  1.3%  1% 1 sig. fig.
3.06
 Note: To avoid round-off error, keep one digit beyond the last
significant figure in all calculations.
- drop only when the final answer is obtained

Round-off errors
Experimental Error & Data Handling
Errors

5.) Propagation of Uncertainty


(i) Multiplication and Division
 The relative uncertainties are used for all numbers in the calculation

Re l . Uncert .Answer  Re l . Uncert .


value1   Re l . Uncert .
2
value 2 2

 Example:

1.76  0.03   1.89  0.02 


 5.64 3 sig. fig.
0.59  0.02 

Re l . Uncert . 
 0.03 ( 100 ),  0.02  ( 100 ) ,  0.02  ( 100 )
1.76 1.89 0.59

Re l . Uncert .  1.7%,  1.1% ,  3.4%

Re l . Uncert .Answer  1.7 2  1.12  3.4 2  4.0%  4% 1 sig. fig.


Experimental Error & Data Handling
Errors

5.) Propagation of Uncertainty


(ii) Once the relative uncertainty of the answer has been obtained, the absolute
uncertainty can also be calculated:

Absolute Uncertaint y
Relative Uncertaint y(%)  ( 100 )
Calculated Value
Rearrange:

Relative Uncertaint y(%)


Absolute Uncertaint y  (calculate d value)
( 100 )

(iii) Example (using the previous example):

 4.0% 
Absolute Uncertaint y  ( 5.64 )   0.23  0.2 1 sig. fig.
 100 
Experimental Error & Data Handling
Errors

5.) Propagation of Uncertainty


(iv) For calculations involving Both additions/subtractions and
multiplication/divisions:
 Treat calculation as a series of individual steps
 Calculate the answer and its uncertainty for each step
 Use the answers and its uncertainty for the next calculation, etc.
 Continue until the final result is obtained

(v) Example:
1.76  0.03   0.59 0.02   0.619 ?  3 sig. fig.
1.89 0.02 
First operation: differences in brackets

1.76  0.03  0.59 0.02   1.17  0.036  3 sig. fig.

0.036  0.032  0.02 2 1 sig. fig., but carry two sig.


fig. through calculation
Experimental Error & Data Handling

Errors

5.) Propagation of Uncertainty


(v) Example:
Second operation: Division

1.17  0.036  1.17  3.1% Convert to relative


  0.61% 3%

1.89  0.02  
1.89  1.1% uncertainty

3 sig. fig.
3.3%  3%  3.1%  1.1%
2 2

1 sig. fig.
Experimental Error & Data Handling

Errors

5.) Propagation of Uncertainty


(vi) Uncertainty of a result should be consistent with the number of significant
figures used to express the result.

(vii) Example:

1.019 (±0.002)
Result & uncertainty match
in decimal place
28.42 (±0.05)

But:
12.532 (±0.064)  too many significant figures

12.53 (±0.06)  reduce to 1 sig. fig. in uncertainty


same reduction in results

The first digit in the answer with any uncertainty associated


with it should be the last significant figure in the number.
Experimental Error & Data Handling

Errors

5.) Common Mistake


(vi) Number of Significant Figures is Not the number shown on your calculator.

Not 10 sig. fig.

23.97
 2.596966414
9.23
Experimental Error & Data Handling

Errors

Example
Find the absolute and percent relative uncertainty and express the answer with
a reasonable number of significant figures:

[4.97 ± 0.05 – 1.86 ± 0.01]/21.1 ± 0.2 =

Das könnte Ihnen auch gefallen